Tuning molecular dynamics by hydration and confinement: antiplasticizing effect of water in hydrated prilocaine nanoclusters.

In glass-forming substances, the addition of water tends to produce the effect of lowering the glass transition temperature, Tg. In a previous work by some of us (Ruiz et al., Sci. Rep., 2017, 7, 7470) we reported on a rare anti-plasticizing effect of water on the molecular dynamics of a simple molecular system, the pharmaceutically active prilocaine molecule, for which the addition of water leads to an increase of Tg. In the present work, we study pure and hydrated prilocaine confined in 0.5 nm and 1 nm pore size molecular sieves, and carry out a comparison with the bulk compounds in order to gain a better understanding of the microscopic mechanisms that result in this rare effect. We find that the Tg of the drug under nanometric confinement can be lower than the bulk value by as much as 17 K. Through the concurrent use of differential scanning calorimetry and broadband dielectric spectroscopy we are able to observe the antiplasticizing effect of water in prilocaine also under nanometric confinement, finding an increase of Tg of up to almost 6 K upon hydration. The extension of our analysis to nanoconfined systems provides a plausible explanation for the very uncommon antiplasticizing effect, based on the formation of water-prilocaine molecular complexes. Moreover, this study deepens the understanding of the behavior of drugs under confinement, which is of relevance not only from a fundamental point of view, but also for practical applications such as drug delivery.

Membrane-containing virus particles exhibit the mechanics of a composite material for genome protection.

The protection of the viral genome during extracellular transport is an absolute requirement for virus survival and replication. In addition to the almost universal proteinaceous capsids, certain viruses add a membrane layer that encloses their double-stranded (ds) DNA genome within the protein shell. Using the membrane-containing enterobacterial virus PRD1 as a prototype, and a combination of nanoindentation assays by atomic force microscopy and finite element modelling, we show that PRD1 provides a greater stability against mechanical stress than that achieved by the majority of dsDNA icosahedral viruses that lack a membrane. We propose that the combination of a stiff and brittle proteinaceous shell coupled with a soft and compliant membrane vesicle yields a tough composite nanomaterial well-suited to protect the viral DNA during extracellular transport.

Broadband nanodielectric spectroscopy by means of amplitude modulation electrostatic force microscopy (AM-EFM).

In this work we present a new AFM based approach to measure the local dielectric response of polymer films at the nanoscale by means of Amplitude Modulation Electrostatic Force Microscopy (AM-EFM). The proposed experimental method is based on the measurement of the tip-sample force via the detection of the second harmonic component of the photosensor signal by means of a lock-in amplifier. This approach allows reaching unprecedented broad frequency range (2-3 × 10(4)Hz) without restrictions on the sample environment. The method was tested on different poly(vinyl acetate) (PVAc) films at several temperatures. Simple analytical models for describing the electric tip-sample interaction semi-quantitatively account for the dependence of the measured local dielectric response on samples with different thicknesses and at several tip-sample distances.

Numerical study of the lateral resolution in electrostatic force microscopy for dielectric samples.

We present a study of the lateral resolution in electrostatic force microscopy for dielectric samples in both force and gradient modes. Whereas previous studies have reported expressions for metallic surfaces having potential heterogeneities (Kelvin probe force microscopy), in this work we take into account the presence of a dielectric medium. We introduce a definition of the lateral resolution based on the force due to a test particle being either a point charge or a polarizable particle on the dielectric surface. The behaviour has been studied over a wide range of typical experimental parameters: tip-sample distance (1-20) nm, sample thickness (0-5) µm and dielectric constant (1-20), using the numerical simulation of the equivalent charge method. For potential heterogeneities on metallic surfaces expressions are in agreement with the bibliography. The lateral resolution of samples having a dielectric constant of more than 10 tends to metallic behaviour. We found a characteristic thickness of 100 nm, above which the lateral resolution measured on the dielectric surface is close to that of an infinite medium. As previously reported, the lateral resolution is better in the gradient mode than in the force mode. Finally, we showed that for the same experimental conditions, the lateral resolution is better for a polarizable particle than for a charge, i.e. dielectric heterogeneities should always look 'sharper' (better resolved) than inhomogeneous charge distributions. This fact should be taken into account when interpreting images of heterogeneous samples.

Positron annihilation and relaxation dynamics from dielectric spectroscopy and nuclear magnetic resonance: cis-trans-1,4-poly(butadiene).

We report a joint analysis of positron annihilation lifetime spectroscopy (PALS), dielectric spectroscopy (BDS), and nuclear magnetic resonance (NMR) on cis-trans-1,4-poly(butadiene) (c-t-1,4-PBD). Phenomenological analysis of the orthopositronium lifetime τ(3)-T dependence by linear fitting reveals four characteristic PALS temperatures: T(b1)(G)=0.63T(g)(PALS), T(g)(PALS), T(b1)(L)=1.22T(g)(PALS), and T(b2)(L)=1.52T(g)(PALS). Slight bend effects in the glassy and supercooled liquid states are related to the fast or slow secondary ß process, from neutron scattering, respectively, the latter being connected with the trans-isomers. In addition, the first bend effect in the supercooled liquid coincides with a deviation of the slow effective secondary ß(eff) relaxation related to the cis-isomers from low-T Arrhenius behavior to non-Arrhenius one and correlates with the onset of the primary α process from BDS. The second plateau effect in the liquid state occurs when τ(3) becomes commensurable with the structural relaxation time τ(α)(T(b2)). It is also approximately related to its crossover from non-Arrhenius to Arrhenius regime in the combined BDS and NMR data. Finally, the combined BDS and NMR structural relaxation data, when analyzed in terms of the two-order parameter (TOP) model, suggest the influence of solidlike domains on both the annihilation behavior and the local and segmental chain mobility in the supercooled liquid. All these findings indicate the influence of the dynamic heterogeneity in both the primary and secondary relaxations due to the cis-trans isomerism in c-t-1,4-PBD and their impact into the PALS response.

Nanodielectric mapping of a model polystyrene-poly(vinyl acetate) blend by electrostatic force microscopy.

We present a simple method to quantitatively image the dielectric permittivity of soft materials at nanoscale using electrostatic force microscopy (EFM) by means of the double pass method. The EFM experiments are based on the measurement of the frequency shifts of the oscillating tip biased at two different voltages. A numerical treatment based on the equivalent charge method allows extracting the values of the dielectric permittivity at each image point. This method can be applied with no restrictions of film thickness and tip radius. This method has been applied to image the morphology and the nanodielectric properties of a model polymer blend of polystyrene and poly(vinyl acetate).

Nanoscale dielectric properties of insulating thin films: from single point measurements to quantitative images.

Dielectric relaxation (DR) has shown to be a very useful technique to study dielectric materials like polymers and other glass formers, giving valuable information about the molecular dynamics of the system at different length and time scales. However, the standard DR techniques have a fundamental limitation: they have no spatial resolution. This is of course not a problem when homogeneous and non-structured systems are analyzed but it becomes an important limitation for studying the local properties of heterogeneous and/or nano-structured materials. To overcome this constrain we have developed a novel approach that allows quantitatively measuring the local dielectric permittivity of thin films at the nanoscale by means of Electrostatic Force Microscopy. The proposed experimental method is based on the detection of the local electric force gradient at different values of the tip-sample distance. The value of the dielectric permittivity is then calculated by fitting the experimental points using the Equivalent Charge Method. Even more interesting, we show how this approach can be extended in order to obtain quantitative dielectric images of insulating thin films with an excellent lateral resolution.

Water dynamics in n-propylene glycol aqueous solutions.

The relaxation dynamics of dipropylene glycol and tripropylene glycol (nPG-n=2,3) water solutions on the nPG-rich side has been studied by broadband dielectric spectroscopy and differential scanning calorimetry in the temperature range of 130-280 K. Two relaxation processes are observed for all the hydration levels; the slower process (I) is related to the alpha relaxation of the solution whereas the faster one (II) is associated with the reorientation of water molecules in the mixture. Dielectric data for process (II) at temperatures between 150 and 200 K indicate the existence of a critical water concentration (x(c)) below which water mobility is highly restricted. Below x(c), nPG-water domains drive the dielectric signal whereas above x(c), water-water domains dominate the dielectric response at low temperatures. The results also show that process (II) at low temperatures is due to local motions of water molecules in the glassy frozen matrix. Additionally, we will show that the glass transition temperatures (T(g)) for aqueous PG, 2PG, and 3PG solutions do not extrapolate to approximately 136 K, regardless of the extrapolation method. Instead, we find that the extrapolated T(g) value for water from these solutions lies in the neighborhood of 165 K.

Combining configurational entropy and self-concentration to describe the component dynamics in miscible polymer blends.

We provide a new approach to describe the component segmental dynamics of miscible polymer blends combining the concept of chain connectivity, expressed in terms of the self-concentration, and the Adam-Gibbs model. The results show an excellent agreement between the prediction of our approach and the experimental data. The self-concentrations obtained yield length scales between 1 and 3.2 nm depending on the temperature, the flexibility of the polymer, expressed in terms of the Kuhn segment, and its concentration in the blends, at temperatures above the glass transition range of the blend.

Dynamics of propylene glycol and its oligomers confined in clay.

The dynamics of propylene glycol (PG) and its oligomers 7-PG and PPG, with Mw = 4000 (about 70 monomers), confined in a Na-vermiculite clay have been investigated by quasi-elastic neutron scattering and dielectric spectroscopy. The liquids are confined to a single molecular layer between the clay platelets, thus giving a true 2D liquid. The results show that the average relaxation time [tau], deduced from neutron scattering at a momentum transfer Q of about 1 A(-1) is in perfect agreement with the dielectric alpha-relaxation time, although neutron scattering does not only probe the main (alpha-) relaxation, but all motions of hydrogens on the experimental time scale. At room temperature 1/[tau] is proportional to Q(2), indicating that the relaxations are mainly due to ordinary translational diffusion. The most unexpected finding is that [tau](or the dielectric alpha-relaxation time) is almost unaffected by the 2D confinement, in contrast to the dielectrically active normal mode of PPG which is substantially slower in the confinement. Only the 7-mer has a significantly slower segmental translational diffusion in the clay. The results suggest that the interactions to the clay surfaces are weak and that the present 2D confinement has a very small influence on the time scale of all our observed relaxation processes, except the normal-mode relaxation.

Dielectric relaxation studies of poly(propylene glycol) confined in vermiculite clay.

The molecular dynamics of oligomeric poly(propylene glycol) (PPG) liquids (M(w)=1200, 2000 and 4000 g/mol) confined in a two-dimensional layer-structured Na-vermiculite clay has been studied by broadband dielectric spectroscopy. In addition to the alpha-relaxation, the normal mode relaxation process was studied for all samples both in bulk and confinement. For the normal mode process the relaxation rate in the clay is drastically shifted to lower frequencies compared to that of the bulk material in contrast to the alpha-process whose relaxation time is only slightly affected by the confinement. Also the temperature dependence of the relaxation time for the normal mode process is strongly affected by the confinement. Moreover, in the clay the intensity of the normal mode is stronger than that of the alpha-process, in contrast to the bulk samples where the opposite is observed.

Post-translational phosphorylation of the slow/beta myosin heavy chain isoform in adult rabbit masseter muscle.

Four different phenotypes of slow muscle fibers, characterized by differential epitope expression in the slow/beta myosin heavy chain (MyHC) isoform, have been identified in adult rabbit masseter muscle. We investigated the role of post-translational phosphorylation in the expression of these four phenotypes. Serial cryostat sections were treated either with alkaline phosphatase to dephosphorylate proteins in the tissue, or with a brain kinase solution and ATP to phosphorylate them, and then stained, using four antibodies that bind specifically to the slow/beta MyHC isoform. In sections pre-treated with phosphatase, immunoreactivity to antibody A4.840 was abolished, but it could be restored by subsequent kinase/ATP treatment or ATP alone, indicating that the expression of its epitope requires phosphorylation. Phosphatase treatment resulted in an exposure of the epitope for antibody A4.951 in cells that normally bind this antibody only weakly or not at all, but since heat treatment alone produced similar effects, the role of phosphorylation in this enhancement is less certain. Immunoreactivity to antibodies S58 and BA-D5 were not influenced by phosphatase pre-treatment. Kinase/ATP treatment was only effective in changing antibody binding when tissues already had been phosphatase treated. We interpret these results to mean that sites of potential phosphorylation may already be occupied by O-linked glycosylation.

A digital 3-dimensional method for computing great artery flows: in vitro validation studies.

BACKGROUND: Conventional 2-dimensional Doppler large vessels are prone to inaccuracy. Three-dimensional (3D) volume imaging provides the opportunity to make cross-sectional flow calculations through digital spatiotemporal integration of flow velocity, area, and profile. METHODS: A new digital 3D color Doppler reconstruction method was used to generate radially acquired flow data sets. Raw scanline data with digital velocity assignments, obtained by scanning parallel to flow, were transferred from a specially programmed but otherwise conventional ultrasonographic system, which controlled a multiplane transesophageal probe, to a computer workstation via an Ethernet link for assimilation into color 3D data sets. This configuration was used to study 20 pulsatile laminar flows (stroke volumes 30 to 70 mL and peak flow rates 65 to 205 mL/s) in a curved tube model with an oval cross-sectional geometry. After generation of the color 3D data set, flow velocity values from cross sections perpendicular to the tubes were analyzed to determine flow rate and stroke volume. RESULTS: The flows from 3D digital velocity profiles showed close correlation with peak instantaneous flow rates (r = 0.99, y = 1.01x-0.9, standard error of estimate 4.1 mL/s). When interpreted with pulsed wave Doppler data obtained through the cardiac cycle, they also allowed computation of stroke volume (r = 0.98, y = 1.44x-2.5, standard error of estimate 3.8 mL). CONCLUSION: The ability to compute laminar flows from 3D digital data sets obtained parallel to the direction of flow and without the need for geometric assumptions represents an important opportunity for and advantage of 3D color Doppler echocardiography.

Sexually dimorphic expression of myosin heavy chains in the adult mouse masseter.

Little is known regarding the role of androgenic hormones in the maintenance of myosin heavy chain (MHC) composition of rodent masticatory muscles. Because the masseter is the principal jaw closer in rodents, we felt it was important to characterize the influence of androgenic hormones on the MHC composition of the masseter. To determine the extent of sexual dimorphism in the phenotype of masseter muscle fibers of adult (10-mo-old) C57 mice, we stained tissue sections with antibodies specific to type IIa and IIb MHC isoforms. Females contain twice as many fibers containing the IIa MHC as males, and males contain twice as many fibers containing the IIb MHC as females. There is a modest amount of regionalization of MHC phenotypes in the mouse masseter. The rostral portions of the masseter are composed mostly of type IIa fibers, whereas the midsuperficial and caudal regions contain mostly type IIb fibers. Using immunoblots, we showed that castration results in an increase in the expression of type IIa MHC fibers in males. Ovariectomy has no effect on the fiber type composition in females. We conclude that testosterone plays a role in the maintenance of MHC expression in the adult male mouse masseter.

200 kD neurofilament protein and synapse elimination in the rat soleus muscle.

We studied the distribution and appearance of the phosphorylated form of the 200 kD neurofilament protein in the rat soleus muscle during the period of postnatal synapse elimination. Unlike many muscles, the appearance of singly innervated muscle cells in soleus occurs well after myogenesis has been completed, so that synapses are eliminated from a stable population of muscle cells. Immunoreactivity to the 200 kD neurofilament protein is present in the terminals of neuromuscular synapses of animals at all postnatal ages from 0 to 21 days. Before postnatal day 10, when physiological studies indicate that all soleus muscle cells receive more than one synaptic input, as many as 30% of soleus muscle cells contain phosphorylated 200 kD neurofilament protein immunoreactivity in only one synaptic terminal. At older ages the number of polyneuronally innervated muscle cells observed using immunostaining is similar to that observed physiologically. These findings suggest that not all developing neuromuscular synapses contain phosphorylated 200 kD neurofilament protein, and that those terminals lacking it comprise most of those eliminated early in the postnatal period. We conclude that the presence of phosphorylated 200 kD neurofilament protein might be highly correlated with the survival of motor nerve terminals during postnatal neuromuscular synapse elimination.

Tenotomy delays both synapse elimination and myogenesis in rat lateral gastrocnemius.

To investigate a possible relationship between synapse elimination and myogenesis, we examined both phenomena during the first 2 weeks of postnatal life in the rat lateral gastrocnemius muscle. Synapse elimination and myogenesis occur simultaneously. Sixty per cent of the number of fibers observed in adult muscles is generated during the first 10 days of postnatal life; during this time, the majority of muscle cells in lateral gastrocnemius also become singly innervated. We delayed synapse elimination by cutting the tendon of insertion of lateral gastrocnemius (tenotomy) on the day of birth. Both synapse elimination and postnatal myogenesis were slowed by tenotomy. Tenotomized muscles contained fewer detectable cells than unoperated contralateral control muscles. These results suggest that synapse elimination may be altered by altering postnatal muscle fiber addition.

Minimal risk of Rh sensitization in induced abortions.

PIP: Evidence from almost 700 consecutive first trimester abortions and 59 random second trimester abortions leads the author to question the need for administering full doses of anti-D globulin to legal abortion patients of Rho-negative blood type routinely. Of 699 first trimester cases, only 17 tested positive for fetal cells. All 17 had less than 2 ml of bleeding, and the remaining cases had no transplacental bleeding; these cases were aborted by curettage. A higher percentage (10%) aborted by saline tested positive, but as in the vacuum curettage group, all bled less than 2 ml. A 20-unit dose, vs. the standard 300-unit dose, is recommended.^ieng

2-methoxyethanol as a solvent for conductometric acid-base titrations.

Conductometric titrations of a series of organic acids in 2-methoxyethanol were performed with 1,3-diphenylguanidine (DPG) as titrant. For benzoic, p-chlorobenzoic and picric acids, phenylphenol and barbital, excellent recoveries were obtained from well-defined conductance vs. volume plots. Results for the dicarboxylic phthalic and adipic acids were also good. However, the titration curves for the difunctional salicylic and p-hydroxybenzoic acids were not clearly defined. The results are discussed and compared with cnductometric titrations of acids in other non-aqueous solvents. Several determinations of electrolyte conductance as a function of concentration revealed that, as expected, the selected substances are weak electrolytes in methoxyethanol.